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Development, optimization and validation of an absolute specific assay for active myeloperoxidase (MPO) and its application in a clinical context: role of MPO specific activity in coronary artery disease

  • Alessandro Trentini ORCID logo EMAIL logo , Valentina Rosta , Savino Spadaro , Tiziana Bellini , Paola Rizzo , Francesco Vieceli Dalla Sega , Angelina Passaro ORCID logo , Giovanni Zuliani , Valentina Gentili , Gianluca Campo and Carlo Cervellati
Published/Copyright: February 7, 2020
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 58 Issue 10

Abstract

Background

Myeloperoxidase (MPO) is an enzyme with a recognized prognostic role in coronary artery disease (CAD), which is also emerging as a promising biomarker for cardiac risk stratification. However, the lack of a consensus method for its quantification has hindered its implementation in clinical practice. The aim of our work was to optimize an absolute sensitive assay for active MPO without external standards, to validate the method in the clinical context of CAD patients, and to estimate the enzyme specific activity.

Methods

In order to determine the MPO concentration using fluorescence readings, this ELISA assay exploits the activity of the enzyme recognized by specific antibodies. The assay was validated in a small cohort of patients that included: healthy subjects (n=60); patients with acute myocardial infarction (AMI, n=25); patients with stable CAD (SCAD, n=25) and a concomitant chronic obstructive pulmonary disease (COPD). Then, total MPO concentration and specific activity (activity/total MPO) were determined.

Results

The assay showed an intra- and inter-assay coefficient of variation of 5.8% and 10.4%, respectively, with a limit of detection (LoD) of 0.074 μU. Both AMI and SCAD patients had higher active and total MPO than controls (p<0.0001 and p<0.01, respectively). The specific activity of MPO was higher in SCAD patients compared to both controls and AMI (p<0.0001).

Conclusions

The study presents a robust and sensitive method for assaying MPO activity in biological fluids with low variability. Moreover, the determination of the specific activity could provide novel insight into the role of MPO in cardiovascular diseases (CVDs).

Keywords: acute myocardial infarction; coronary artery disease; myeloperoxidase; oxidative stress; stable CAD
Corresponding author: Alessandro Trentini, PhD, Department of Biomedical and Specialist Surgical Sciences, Section of Medical Biochemistry, Molecular Biology and Genetics, University of Ferrara, Via Luigi Borsari 46, I-44121 Ferrara, Italy, Phone: +39-532-455441, Fax. +39-532-455426

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: This study was supported by “Fondo di Ateneo per la Ricerca” (FAR) grant number: 2018-FAR.L-CC_006.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2019-0817).

Received: 2019-08-05
Accepted: 2020-01-04
Published Online: 2020-02-07
Published in Print: 2020-09-25

©2020 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2019-0817
Keywords for this article
acute myocardial infarction; coronary artery disease; myeloperoxidase; oxidative stress; stable CAD

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Blood sampling for metanephrines: to stick or stick and wait?
  4. Review
  5. Glycan-specific antibodies as potential cancer biomarkers: a focus on microarray applications
  6. Mini Review
  7. Detection of circulating anti-skin antibodies by indirect immunofluorescence and by ELISA: a comparative systematic review and meta-analysis
  8. Opinion Papers
  9. The shift of the paradigm between ageing and diseases
  10. Managing COVID-19 outbreak in Nigeria: matters arising
  11. Circulating tumor DNA (ctDNA) is not a good proxy for liquid biopsies of tumor tissues for early detection
  12. General Clinical Chemistry and Laboratory Medicine
  13. Exact time of venous blood sample collection – an unresolved issue, on behalf of the European Federation for Clinical Chemistry and Laboratory Medicine (EFLM) Working Group for Preanalytical Phase (WG-PRE)
  14. Percentile transformation and recalibration functions allow harmonization of thyroid-stimulating hormone (TSH) immunoassay results
  15. Interference of anti-streptavidin antibodies in immunoassays: a very rare phenomenon or a more common finding?
  16. Blood sampling for metanephrines comparing venipuncture vs. indwelling intravenous cannula in healthy subjects
  17. Volumetric absorptive microsampling and dried blood spot microsampling vs. conventional venous sampling for tacrolimus trough concentration monitoring
  18. Trueness evaluation and verification of inter-assay agreement of serum folate measuring systems
  19. Effects of endurance exercise on serum concentration of calcitonin gene-related peptide (CGRP): a potential link between exercise intensity and headache
  20. Comprehensive characterization and resolution of discrepant spectrophotometric bilirubin results in patients on eltrombopag therapy
  21. Influence of pancreatic status on circulating plasma sterols in patients with cystic fibrosis
  22. Influence of isotopically labeled internal standards on quantification of serum/plasma 17α-hydroxyprogesterone (17OHP) by liquid chromatography mass spectrometry
  23. Reference Values and Biological Variations
  24. The European Biological Variation Study (EuBIVAS): weekly biological variation of cardiac troponin I estimated by the use of two different high-sensitivity cardiac troponin I assays
  25. Cardiovascular Diseases
  26. Comparison of the diagnostic performance with whole blood and plasma of four rapid antibody tests for SARS-CoV-2
  27. Infectious Diseases
  28. Exploring the possibilities of infrared spectroscopy for urine sediment examination and detection of pathogenic bacteria in urinary tract infections
  29. Letters to the Editors
  30. Towards the rational utilization of SARS-CoV-2 serological tests in clinical practice
  31. Response of anti-SARS-CoV-2 total antibodies to nucleocapsid antigen in COVID-19 patients: a longitudinal study
  32. Development, optimization and validation of an absolute specific assay for active myeloperoxidase (MPO) and its application in a clinical context: role of MPO specific activity in coronary artery disease
  33. The early antibody response to SARS-Cov-2 infection
  34. Laboratory work safety rules and guidelines during COVID-19 pandemic in Polish clinical laboratories – do our laboratories work according to a recent IFCC Taskforce Recommendations?
  35. Serum prealbumin deserves more significance in the early triage of COVID-19 patients
  36. Reference intervals for clinically reportable platelet parameters on the Mindray BC-6800Plus hematology analyzer
  37. A new method for monitoring harmonization of laboratory results within EQA schemes
  38. Potential serum magnesium under request in primary care. Laboratory interventions to identify patients with hypomagnesemia
  39. Interference from immunocomplexes on a high-sensitivity cardiac troponin T immunoassay
  40. Interleukin-6 chemiluminescent immunoassay on Lumipulse G600 II: analytical evaluation and comparison with three other laboratory analyzers
  41. Detection of Hb Phnom Penh by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry during the measurement of glycated hemoglobin
  42. Persistently increased vitamin B12 concentration due to cobalamin macrocomplexes: a case report and review of the literature
  43. Antidepressant use limits serotonin as a marker for neuroendocrine tumor disease activity by lowering of circulating serotonin concentrations
  44. Limitations of rapid diagnostic testing in the work-up of dengue infection – a case report
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